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Originally published as Biophys J. BioFAST on March 11, 2005.
doi:10.1529/biophysj.104.054155
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Biophysical Journal 88:4032-4044 (2005)
© 2005 The Biophysical Society

Building Up of the Liquid-Ordered Phase Formed by Sphingomyelin and Cholesterol

C. Chachaty *, D. Rainteau *, C. Tessier *, P. J. Quinn {dagger} and C. Wolf *

* Universite Paris 6, INSERM U538, Paris 75012, France; and {dagger} Life Sciences, King's College London, SE1 9NH, UK

Correspondence: Address reprint requests to Claude Wolf, E-mail: wolf{at}ccr.jussieu.fr.

The long-range and molecular orders and dynamics in codispersions of egg sphingomyelin-cholesterol have been investigated by synchrotron x-ray diffraction and electron spin resonance using phosphatidylcholine spin-labeled at several positions on the sn-2 chain. Mixtures containing 0, 17, 33, 41, 50 mol% cholesterol exhibited a single phase by x-ray diffraction methods. The temperature dependence of the d-spacing between 20 and 50°C is attenuated with increasing proportions of cholesterol, becoming invariant for cholesterol contents of 41 and 50 mol% on completion of the liquid-ordered phase. Electron spin resonance revealed two sites for 17 and 33 mol% cholesterol. One site is highly ordered and the other is less ordered than the fluid phase of pure sphingomyelin as shown by the molecular and the intramolecular order parameters reflecting the segmental motions of the probe. The two-sites exchange rate indicates a mean lifetime of the sites of ~0.1 µs during which the lipid displacement is ~1 nm. The short lifetime of the sites probed by ESR and the single phase detected by x-ray diffraction support in this binary mixture, the building up of the Lo phase by a progressive accumulation of randomly distributed sphingomyelin-cholesterol condensed complexes rather than by diffusional exchange between extended domains.




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